| Assay Method Information | |
| | In Vitro Cell-Free Assay |
| Description: | In a final reaction volume of 50 μl, CK2 ααββ (4 ng, 8.5 mU) was incubated with various concentrations of test compounds in DMSO (1 ul, 2% by volume), 20 mM MOPS pH 7.2, 10 mM EGTA, 0.15 M NaCl, 10 mM DTT, 0.002% Brij-35, 200 μM RRRDDDSDDD (SEQ ID NO.: 4), 10 mM MgAcetate, ATP 15 uM and 0.33% (by volume) (hr-3311ATP: Stock 1 mCi/100 μl; 3000 Ci/mmol (Perkin Elmer)). Reactions were maintained for 40 min at 23° C. The reactions were quenched with 100 ul of 0.75% Phosphoric acid, then transferred to and filtered through a Phosphocellulose filter plate (Millipore, MSPH-N6B-50). After washing each well 4 times with 0.75% Phosphoric acid, scintillation fluid (20 uL) was added to each well and the residual radioactivity was measured using a Wallac luminescence counter. |
| Affinity data for this assay | |
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